To stain TMRM in the whole brain of adult Drosophila, I referenced several methods, ultimately following the method below:
1. Dissect 3 adult brains in cold Drosophila Schneider media (DSM)
2. Incubate in 200nM TMRM with DSM - for 40min at RT
3. Mount samples using 50nM TMRM with DSM for Imaging
However, I could not achieve the staining quality like that of the attached image.
So, I changed the method and tried out various experimental conditions:
I tried TMRM concentration of 400nM and 600nM, TMRM incubation temperature of 4℃, and TMRM incubation time of 15min and 1hour.
But, the results were the same.
How can I solve this problem?